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Molecular cloning and expression analysis of a monosaccharide transporter gene OsMST4 from rice (Oryza sativa L.)

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Author: Wang, Y. · Xu, H. · Wei, X. · Chai, C. · Xiao, Y. · Zhang, Y. · Chen, B. · Xiao, G. · Ouwerkerk, P.B.F. · Wang, M. · Zhu, Z.
Type:article
Date:2007
Institution: TNO Kwaliteit van Leven
Source:Plant Molecular Biology, 4, 65, 439-451
Identifier: 240257
doi: doi:10.1007/s11103-007-9228-x
Keywords: Biology · Biomedical Research · cDNA cloning · Gene expression · Monosaccharide transporter · Oryza sativa L. · Seed development · Cloning · Crops · Fructose · Genes · Physiology · Plant cell culture · Transcription · cDNA cloning · Monosaccharide transporter · Oryza sativa L. · Seed development · Molecular biology · complementary DNA · glucose transporter · messenger RNA · primer DNA · vegetable protein · amino acid sequence · article · chemistry · gene expression profiling · genetics · in situ hybridization · molecular cloning · molecular genetics · nucleotide sequence · phylogeny · reverse transcription polymerase chain reaction · rice · sequence homology · Amino Acid Sequence · Base Sequence · Cloning, Molecular · DNA Primers · DNA, Complementary · Gene Expression Profiling · In Situ Hybridization · Molecular Sequence Data · Monosaccharide Transport Proteins · Oryza sativa · Phylogeny · Plant Proteins · Reverse Transcriptase Polymerase Chain Reaction · RNA, Messenger · Sequence Homology, Amino Acid · Oryza sativa · Saccharomyces cerevisiae

Abstract

Monosaccharide transporters mediate the membrane transport of a variable range of monosaccharides, which plays a crucial role in sugar distribution throughout the plant. To investigate the significance of monosaccharide transporters for rice (Oryza sativa L.) seed development, cDNA of a new putative monosaccharide transporter gene OsMST4 was isolated. The deduced OsMST4 protein shows typical features of monosaccharide transporters, and shares high homology with other plant homologues. Heterologous expression in yeast (Saccharomyces cerevisiae) showed that OsMST4 is a functional monosaccharide transporter capable of transporting glucose, fructose, mannose and galactose. Transcriptional analysis revealed that OsMST4 is expressed in all tested organs/tissues. In developing caryopses, its expression is high at the early and middle grain filling stages, and declines gradually to low levels after that. Further analysis revealed that it is expressed in both the maternal tissue and the filial tissue, with its highest expression in embryo. Cellular location in young caryopses through RNA in situ hybridization showed that OsMST4 mRNA mainly accumulates in the vascular parenchyma of the chalazal vein, cross-cells, nucellar tissue and endosperm. The expression pattern of OsMST4 was further confirmed by histochemical analysis of the OsMST4-promoter-β-glucuronidase (GUS) transgenic rice plants. These data indicate that OsMST4 is actively involved in monosaccharides supply for seed development during the course of grain filling. In addition, the cell type-specific expression patterns of OsMST4 in other sink and source tissues were also investigated, and its corresponding physiological roles were discussed. © 2007 Springer Science+Business Media B.V.